Typical DNA yield and purity from 1.5 ml of bacterial culture (LB media) for different cells and vectors.|The simple bind-wash-elute procedure of the QuickLyse system.|Plasmid DNA (pSV-β-galactosidase) was purified using the QuickLyse Miniprep Kit (QL), as well as three additional commercially available kits (1–3). Purified plasmid DNA was digested with NdeI and EcoRI restriction enzymes. The reactions were performed in duplicate. M: markers. Ø: uncut plasmid.|Typical sequencing results for plasmid DNA purified using the QuickLyse Miniprep Kit. The pSV-β-galactosidase plasmid was isolated from TOP10 cells and sequenced using the BigDye terminator sequencing chemistry.|

The QIAGEN QuickLyse Miniprep Kit uses one-step lysis technology for ultrafast preparation of up to 15 µg plasmid DNA. The quality of the DNA ensures robust performance in applications, such as sequencing, restriction analyses, and cloning (see figure "Reproducible yields").

The QuickLyse Miniprep Kit yields sufficient plasmid DNA for any downstream application, including sequencing (see figure "Long read-lengths in automated sequencing"). Up to 15 µg of high-copy plasmid DNA can be obtained from 1.5 ml of bacterial culture (see figure "High yields of high-quality plasmid DNA"). Isolated plasmid DNA has OD260/280 ratios of 1.7–1.9, indicating that the quality and purity is suitable for all sensitive applications. Sequencing read lengths of over 700 bases are easily achieved.

Principle

Unlike most protocols that use a 3-step lysis procedure, the QuickLyse Miniprep Kit combines enzymatic and osmotic processes to lyse bacterial cells in a single, 3-minute step. In addition, QuickLyse technology uses fewer buffers, simplifying handling and saving time: 24 plasmid DNA minipreps can be prepared in less than 22 minutes.

Procedure

Bacterial cells are resuspended and lysed and DNA is bound in a single buffer, enabling fast DNA purification with minimal hands-on time. After a 3-minute lysis step, the clear lysate is applied directly to a QuickLyse Spin Column. Plasmid DNA binds to the column membrane and is eluted after a single wash step (see flowchart "QuickLyse procedure"). The isolated DNA can be used directly in standard applications, including automated sequencing, PCR, restriction analysis, and cloning.